Brilliant cresyl blue staining allows the selection for developmentally competent immature feline oocytes.

Abstract

The outcome of in-vitro-maturation and in-vitro-fertilization of feline oocytes depends on the selection of high quality oocytes, and is often restricted to morphological criteria. The aim of this study was to test whether the Brilliant cresyl blue (BCB) staining is suitable for pre-selection of feline oocytes before in-vitro-maturation. Cumulus-oocytes-complexes (COC) were released from domestic cat ovaries obtained after ovariectomy and were subjected to BCB staining. BCB+\xa0stained oocytes were characterized by a violet/pale blue staining of the ooplasma, BCB- oocytes remained unstained. Transmission electron microscopy indicated for a slightly advanced stage of BCB- oocytes within the maturation process. After 24\u202fh in-vitro-maturation, almost 75% of BCB+ and 21.5% of BCB- oocytes were able to reach metaphase II. Also, after in-vitro-fertilization, significantly more oocytes developed to morulae (19.2%) if oocytes were preselected for BCB staining, although 8% of unstained COC still reached advanced embryo stages. Prolonged storage of ovaries before COC retrieval for 16-20\xa0h\xa0at 4\xa0°C was accompanied by reduced number of BCB+\xa0oocytes (96 of 210, 45.7%) in comparison to freshly isolated COC (151 of 225, 67.1%), and impaired cleavage rate (19.8%) and morula rate (9.4%) of BCB+\xa0oocytes but the rate of embryos which developed to advanced stages remained unchanged (∼50%). To conclude, BCB staining is a very useful tool to preselect immature COC of feline species ensuring higher developmental rates.